Human B7-H3 Antibody

Name: Human B7-H3 Antibody
Brand: R&D Systems
SKU: AF1027
Rating: 4 (1 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AF1027

Citations (18)

(1)

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Key Product Details

Species Reactivity:	Human
Immunogen:	Mouse myeloma cell line NS0-derived recombinant human B7‑H3 Leu29-Pro245 Accession # NP_079516
Applications:	CyTOF-ready, Flow Cytometry, Immunohistochemistry, Knockout Validated, Simple Western, Western Blot
Label:	Unconjugated
Source:	Polyclonal Goat IgG
Specificity:	Detects human B7-H3 in direct ELISAs and Western blots.

View Full Technical Details for Human B7-H3 Antibody

Applications for Human B7-H3 Antibody

	Recommended Concentration	Sample
Western Blot	1 µg/mL	See below
Simple Western	10 µg/mL	See below
Flow Cytometry	0.25 µg/10⁶ cells	See below
Immunohistochemistry	5-15 µg/mL	See below
CyTOF-ready	Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Item	Value
Knockout Validated	B7‑H3 is specifically detected in U2OS human osteosarcoma parental cell line but is not detectable in B7‑H3 knockout U2OS cell line.

Please Note: Optimal dilutions should be determined by each laboratory for each application.

Scientific Data Examples for Human B7-H3 Antibody

	Western Blot Detection of Human B7‑H3 by Western Blot. Western blot shows lysates of LNCaP human prostate cancer cell line and U2OS human osteosarcoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7-H3 at approximately 90-110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
	Flow Cytometry Detection of B7‑H3 in PC‑3 Human Cell Line by Flow Cytometry. PC-3 human prostate carcinoma cells were stained with Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).
	Immunohistochemistry B7-H3 in Human Melanoma. B7-H3 was detected in immersion fixed paraffin-embedded sections of human melanoma using Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
	Simple Western Detection of Human B7‑H3 by Simple Western^TM. Simple Western lane view shows lysates of U2OS human osteosarcoma cell line, loaded at 0.2 mg/mL. A specific band was detected for B7-H3 at approximately 120-160 kDa (as indicated) using 10 µg/mL of Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
	Knockout Validated Western Blot Shows Human B7‑H3 Specificity by Using Knockout Cell Line. Western blot shows lysates of U2OS human osteosarcoma parental cell line and B7-H3 knockout U2OS cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7-H3 at approximately 95 kDa (as indicated) in the parental U2OS cell line, but is not detectable in knockout U2OS cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Formulation, Preparation and Storage

Purification	Antigen Affinity-purified
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 6 months, -20 to -70 °C under sterile conditions after reconstitution.

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Background: B7-H3

Human B7 homolog 3 (B7-H3) is a member of the B7 family of immune proteins that provide signals for regulating immune responses (1‑3). Other family members include B7-1, B7-2, B7-H2, PD-L1 (B7-H1), and PD-L2. B7 proteins are immunoglobulin (Ig) superfamily members with extracellular Ig-V-like and Ig-C-like domains and short cytoplasmic domains. Among the family members, they share about 20‑40% amino acid (aa) sequence identity. The cloned human B7-H3 cDNA encodes a 316 aa type I membrane precursor protein with a putative 28 aa signal peptide, a 217 aa extracellular region containing one V-like and one C-like Ig domain, a transmembrane region, and a 45 aa cytoplasmic domain. An isoform of human B7-H3 containing a four-Ig-like domain extracellular region has also been identified. Human B7-H3 is not expressed on resting B cells, T cells, monocytes or dendritic cells, but is induced on dendritic cells and monocytes by inflammatory cytokines. B7-H3 expression is also detected on various normal tissues and in some tumor cell lines. Human B7-H3 does not bind any known members of the CD28 family of immunoreceptors. However, B7-H3 has been shown to bind an unidentified counter-receptor on activated T cells to costimulate the proliferation of CD4⁺ or CD8⁺ T cells. B7-H3 has also been found to enhance the induction of primary cytotoxic T lymphocytes and stimulate IFN-gamma production (1‑3).

Item	Value
References	Chapoval, A.I. et al. (2001) Nat. Immunol. 2:269. Sharpe, A.H. and G.J. Freeman (2002) Nat. Rev. Immunol. 2:116. Coyle, A. and J. Gutierrez-Ramos (2001) Nat. Immunol. 2:203.
Long Name	B7 Homolog 3
Entrez Gene IDs	80381 (Human); 102657 (Mouse); 315716 (Rat); 102131295 (Cynomolgus Monkey)
Alternate Names	B7-H3, B7-H3B7 homolog 3, B7H3, B7H34Ig-B7-H3, CD276, CD276 antigen, CD276 molecule, Costimulatory molecule

Citations for Human B7-H3 Antibody (18)

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Citations are publications that use Bio-Techne products. Selected citations for Human B7-H3 Antibody include:

Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

J Joung et al. (2022), CRISPR activation screen identifies BCL-2 proteins and B3GNT2 as drivers of cancer resistance to T cell-mediated cytotoxicity Nature Communications, 13(1):1606.
PMID: 35338135
Species: Human
Sample Types: Whole Tissue
Applications: IHC

R Wang et al. (2021), B7-H3 suppresses doxorubicin-induced senescence-like growth arrest in colorectal cancer through the AKT/TM4SF1/SIRT1 pathway Cell Death Disease, 2021;12(5):453.
PMID: 33958586
Species: Human
Sample Types: Cell Culture Lysates
Applications: Western Blot

Y Ma et al. (2020), B7-H3 promotes the cell cycle-mediated chemoresistance of colorectal cancer cells by regulating CDC25A J Cancer, 11(8):2158-2170.
PMID: 32127943
Y Ma et al. (2020), B7-H3 regulates KIF15-activated ERK1/2 pathway and contributes to radioresistance in colorectal cancer Cell Death Dis, 11(10):824.
PMID: 33011740
Species: Human
Sample Types: Whole Tissue
Applications: IHC

R Cheng et al. (2020), CD276 Promotes Vasculogenic Mimicry Formation in Hepatocellular Carcinoma via the PI3K/AKT/MMPs Pathway Onco Targets Ther, 13(0):11485-11498.
PMID: 33204103
Species: Human
Sample Types: Tissue Homogenates
Applications: Western Blot

B Ganesan et al. (2020), Clinical relevance of B7H3 expression in retinoblastoma Sci Rep, 10(1):10185.
PMID: 32576886
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

J Zhang et al. (2019), B7H3 regulates differentiation and serves as a potential biomarker and theranostic target for human glioblastoma Lab. Invest., 0(0).
PMID: 30914782
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

T Shi et al. (2019), B7-H3 promotes aerobic glycolysis and chemoresistance in colorectal cancer cells by regulating HK2 Cell Death Dis, 10(4):308.
PMID: 30952834
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P

Y Li et al. (2017), B7-H3 Promotes the Migration and Invasion of Human Bladder Cancer Cells via the PI3K/Akt/STAT3 Signaling Pathway J Cancer, 8(5):816-824.
PMID: 28382144
Species: Human
Sample Types: Whole Tissue
Applications: IHC

D Quandt et al. (2017), High constitutive B7-H3 expression on human keratinocytes supports T cell immunity J. Dermatol. Sci., 0(0).
PMID: 28465042
Species: Human
Sample Types: Whole Tissue
Applications: IHC

KE Wilson et al. (2017), Spectroscopic Photoacoustic Molecular Imaging of Breast Cancer using a B7-H3-targeted ICG Contrast Agent Theranostics, 7(6):1463-1476.
PMID: 28529630
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

BR Yoon et al. (2016), Preferential Induction of the T Cell Auxiliary Signaling Molecule B7-H3 on Synovial Monocytes in Rheumatoid Arthritis J. Biol. Chem, 291(8):4048-57.
PMID: 26702052
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P

Bachawal S et al. (2015), Breast Cancer Detection by B7-H3-Targeted Ultrasound Molecular Imaging. Cancer Res, 75(12):2501-9.
PMID: 25899053
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Sun J et al. (2011), Origination of new immunological functions in the costimulatory molecule B7-H3: the role of exon duplication in evolution of the immune system. PLoS ONE, 6(9):e24751.
PMID: 21931843
Species: Human
Sample Types: Whole Cells
Applications: ICC

Tran CN et al. (2008), Interactions of T cells with fibroblast-like synoviocytes: role of the B7 family costimulatory ligand B7-H3. J. Immunol., 180(5):2989-98.
PMID: 18292521
Species: Bovine
Sample Types: Cell Lysates
Applications: Western Blot

Nagai Y et al. (2008), Anterior pituitary progenitor cells express costimulatory molecule 4Ig-B7-H3. J. Immunol., 181(9):6073-81.
PMID: 18941196
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

Jang BC et al. (2007), 12-O-tetradecanoyl phorbol 13-acetate induces the expression of B7-DC, -H1, -H2, and -H3 in K562 cells. Int. J. Oncol., 31(6):1439-47.
PMID: 17982670
Species: Human
Sample Types: Tissue Homogenates
Applications: Western Blot

Petroff MG et al. (2005), The immunomodulatory proteins B7-DC, B7-H2, and B7-H3 are differentially expressed across gestation in the human placenta. Am. J. Pathol., 167(2):465-73.
PMID: 16049332

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Human B7-H3 Antibody

Name: Anonymous

Application: Simple Western

Sample Tested: b7h3 4ig protein,Purified protein

Species: Human

Verified Customer | Posted 11/02/2018

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Human B7-H3 Antibody

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

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Applications for Human B7-H3 Antibody

Scientific Data Examples for Human B7-H3 Antibody

Western Blot

Flow Cytometry

Immunohistochemistry

Simple Western

Knockout Validated

Formulation, Preparation and Storage

Reconstitution Calculator

Background: B7-H3

Citations for Human B7-H3 Antibody (18)

Customer Reviews for Human B7-H3 Antibody (1)

Customer Images

Human B7-H3 Antibody

Product Documents for Human B7-H3 Antibody

Product Datasheets

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